Jia Peipei, Li Jingyi, Wu Jilin, Li Xueying, Mao Sicong, Wang Sainan, Dong Yanmei
National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Department of Cariology and Endodontology, National Clinical Research Center for Oral Diseases, Peking University School and Hospital of Stomatology, Beijing, China.
National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Fourth Division, Department of General Dentistry, National Clinical Research Center for Oral Diseases, Peking University School and Hospital of Stomatology, Beijing, China.
J Endod. 2025 Jun 6. doi: 10.1016/j.joen.2025.06.002.
To investigate the effects of dentin matrix-extracted complex proteins (DMEPs), especially basic-extracted DMEP (bDMEP), on the regulation of pulp inflammation and repair processes.
Protein components of neutral-extracted DMEP (nDMEP) and bDMEP were analyzed using a cytokine array and ELISA. Human dental pulp cells stimulated by lipopolysaccharide (LPS-hDPCs) were treated with nDMEP and bDMEP. CCK-8 and transwell experiments were employed to assess cell proliferation and migration. The expression levels of genes and proteins related to inflammation and odontogenic differentiation were analyzed through quantitative real-time polymerase chain reaction, ELISA, immunofluorescence staining, and western blot. In vivo, gelatin sponges containing nDMEP or bDMEP were employed as pulp-capping materials in a rat pulpitis model. Histological staining was performed to evaluate inflammation and tissue repair in inflamed pulp tissue.
Compared with nDMEP, alkaline treatment significantly increased the content of multiple growth factors (C-X-C motif chemokine ligand 14, bone morphogenetic protein-2, transforming growth factor-beta1, transforming growth factor-beta2, osteopontin, insulin-like growth factor binding protein-2) and decreased some inflammatory mediators (macrophage inflammatory protein-1, interleukin (IL)-17, intercellular adhesion molecule-2, neural cell adhesion molecule-1) in DMEP. bDMEP enhanced the proliferation and chemotaxis of LPS-hDPCs. DMEP, especially bDMEP, inhibited the levels of proinflammatory cytokines (IL-1β, IL-6, tumor necrosis factor-α, CXCL10) while enhancing the expression of odontogenesis-related genes (dentin matrix protein 1, dentin sialophosphoprotein, collagen type I, alkaline phosphatase) in LPS-hDPCs. In vivo, bDMEP effectively reduced inflammatory infiltration and facilitated formation of dentin-like mineralization in a rat pulpitis model.
Alkaline treatment significantly improved the bioactivity of growth factors derived from the dentin matrix, alleviating the pulpal inflammatory response and facilitating biomimetic repair in inflamed pulp tissue.
研究牙本质基质提取的复合蛋白（DMEPs），尤其是碱提取的DMEP（bDMEP）对牙髓炎症调节和修复过程的影响。
使用细胞因子阵列和酶联免疫吸附测定（ELISA）分析中性提取的DMEP（nDMEP）和bDMEP的蛋白质成分。用nDMEP和bDMEP处理经脂多糖刺激的人牙髓细胞（LPS-hDPCs）。采用细胞计数试剂盒-8（CCK-8）和Transwell实验评估细胞增殖和迁移。通过定量实时聚合酶链反应、ELISA、免疫荧光染色和蛋白质免疫印迹分析与炎症和牙源性分化相关的基因和蛋白质表达水平。在体内，将含有nDMEP或bDMEP的明胶海绵用作大鼠牙髓炎模型中的盖髓材料。进行组织学染色以评估炎症牙髓组织中的炎症和组织修复情况。
与nDMEP相比，碱处理显著增加了DMEP中多种生长因子（C-X-C基序趋化因子配体14、骨形态发生蛋白-2、转化生长因子-β1、转化生长因子-β2、骨桥蛋白、胰岛素样生长因子结合蛋白-2）的含量，并降低了一些炎症介质（巨噬细胞炎性蛋白-1、白细胞介素（IL）-17、细胞间黏附分子-2、神经细胞黏附分子-1）的含量。bDMEP增强了LPS-hDPCs的增殖和趋化性。DMEP，尤其是bDMEP，在LPS-hDPCs中抑制促炎细胞因子（IL-1β、IL-6、肿瘤坏死因子-α、CXCL10）水平，同时增强牙源性分化相关基因（牙本质基质蛋白1、牙本质涎磷蛋白、I型胶原、碱性磷酸酶）的表达。在体内，bDMEP在大鼠牙髓炎模型中有效减少炎症浸润并促进牙本质样矿化形成。
碱处理显著提高了牙本质基质衍生生长因子的生物活性，减轻了牙髓炎症反应，促进了炎症牙髓组织的仿生修复。
